Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/20198
DC FieldValueLanguage
dc.contributor許美鈴zh_TW
dc.contributorMeei-Ling Sheuen_US
dc.contributor.author賴德偉zh_TW
dc.contributor.authorLai, De-Weien_US
dc.contributor.other生物醫學研究所zh_TW
dc.date2013en_US
dc.date.accessioned2014-06-06T07:12:06Z-
dc.date.available2014-06-06T07:12:06Z-
dc.identifierU0005-0107201314491000en_US
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The epithelial-mesenchymal transition (EMT) phenomenon. Annals of Oncology 21, vii89-vii92. 36. Kallergi, G., Papadaki, M.A., Politaki, E., Mavroudis, D., Georgoulias, V., and Agelaki, S. (2011). Epithelial to mesenchymal transition markers expressed in circulating tumour cells of early and metastatic breast cancer patients. Breast Cancer Research 13, R59. 37. Amparo Cano, M.A.P.-M., Isabel Rodrigo, Annamaria Locascio, Maria J. Blanco,, and Marta G. del Barrio, F.P.a.M.A.N. (2000). The Transcription Factor Snail Controls Epithelial–Mesenchymal Transitions by Repressing E-cadherin Expression. Journal of Clinical Investigation 2, 76-83. 38. Takkunen, M., Grenman, R., Hukkanen, M., Korhonen, M., Garcia de Herreros, A., and Virtanen, I. (2006). Snail-dependent and -independent Epithelial-Mesenchymal Transition in Oral Squamous Carcinoma Cells. Journal of Histochemistry and Cytochemistry 54, 1263-1275. 39. Hui Li, H.W., Fang Wang, Qing Gu, Xun Xu (2011). Snail Involves in the Transforming Growth Factor b1-Mediated Epithelial-Mesenchymal Transition of Retinal Pigment Epithelial Cells. Journal of Clinical Investigation 6, 1-9. 40. Kang, Y., and Massague, J. (2004). Epithelial-Mesenchymal Transitions. Cell 118, 277-279. 41. Antonio Garcia De Herreros , S.P., Mayssaa Nassour , Pierre Savagner (2012). Snail family regulation and epithelial mesenchymal transitions in breast cancer 42. Guaita, S. (2002). Snail Induction of Epithelial to Mesenchymal Transition in Tumor Cells Is Accompanied by MUC1 Repression and ZEB1 Expression. Journal of Biological Chemistry 277, 39209-39216. 43. Fendrich, V., Waldmann, J., Feldmann, G., Schlosser, K., Konig, A., Ramaswamy, A., Bartsch, D.K., and Karakas, E. (2008). Unique expression pattern of the EMT markers Snail, Twist and E-cadherin in benign and malignant parathyroid neoplasia. European Journal of Endocrinology 160, 695-703. 44. Moreno-Bueno, G., Cubillo, E., Sarrio, D., Peinado, H., Rodriguez-Pinilla, S.M., Villa, S., Bolos, V., Jorda, M., Fabra, A., Portillo, F., et al. (2006). Genetic Profiling of Epithelial Cells Expressing E-Cadherin Repressors Reveals a Distinct Role for Snail, Slug, and E47 Factors in Epithelial-Mesenchymal Transition. Cancer Research 66, 9543-9556. 45. Myong, N.-H. (2012). Loss of E-cadherin and Acquisition of Vimentin in Epithelial-Mesenchymal Transition are Noble Indicators of Uterine Cervix Cancer Progression. Korean Journal of Pathology 46, 341.cancer progression. J Mammary Gland Biol Neoplasia 294, 171-221. 46. Lee, K., and Nelson, C.M. (2012). New Insights into the Regulation of Epithelial–Mesenchymal Transition and Tissue Fibrosis. International Review of Cell and Molecular Biology 294, 173-206.en_US
dc.identifier.urihttp://hdl.handle.net/11455/20198-
dc.description.abstract丁香酚(4-allyl-2-methoxyphenol)已經知道具有抗發炎的功用, 但是在抑制癌症以及抑制腹膜轉移的機制尚未清楚。在本研究中,我們的目的是評估在體內和體外的抗腫瘤生長和抗轉移的潛力。我們的結果發現,經由丁香酚處理原位植入MKN45胃癌細胞的小鼠不管在腫瘤的生長以及腹膜傳播和肝/肺轉移有顯著減少,並且伴隨著誘導細胞凋亡。此外,在給予丁香酚後,在胃癌細胞中發現相對增加內質網壓力的標記蛋白,GADD153,IRE1a,P-PERK,P-elf2a,Caspase7等蛋白表達。另外芳香烴受體(AHR)和COX-2是環境致癌物的活化轉錄因子與腫瘤發生和轉移因子。同時,丁香酚處理也減少了AhR/RelA相互作用和COX-2 基因的表達。如同上述,我們使用AGS SCM-1和N87胃癌細胞在體外處理皆可發現相同結果。因此進一步在細胞凋亡的部分我們發現在給予丁香酚後,胃癌細胞增加內質網壓力的標記蛋白GADD153和死亡受體5主要是經由增加(CHOP)/ DR5上游啟動子結合活性,但不是死亡受體4。此外,丁香酚處理後的胃癌細胞中AhR的降解和在內質網產生壓力有相關聯性,如同上述內質網產生壓力時相對產生特定鈣蛋白酶-10和AhR的相互作用增加,促使將AhR蛋白的降解,但不是鈣蛋白酶-1或鈣蛋白酶-2。沉默鈣蛋白酶-10可以發現相對應的抑制由丁香酚治療的生物效應。而不管在隨著時間或濃度劑量效應都可以發現丁香酚處理後AhR/RelA相互作用減少也會相對影響核易位和在腫瘤細胞中的DNA結合活性並且活化鈣蛋白酶-10將AhR降解。兩者合計,我們的研究結果表明,丁香酚抑制胃腫瘤的生長和腹膜轉移是經由激活內質網壓力和誘導細胞凋亡。zh_TW
dc.description.abstractEugenol (4-allyl-2-methoxyphenol) is known to suppress the inflammation; however, its anti-tumor growth, anti-peritoneal dissemination effects have not been studied so far in orthotopic mouse model. In the present study, we aimed to evaluate the anti-tumor growth and anti-metastatic potential of Eugenol in vivo and in vitro. Our results demonstrate that tumor growth, peritoneal dissemination and liver/lung metastasis of orthotopically implanted MKN45 cells were significantly reduced in Eugenol-treated mice along with the induction of apoptosis. Furthermore, Eugenol-treated tumors showed increased ER stress signature such as increased expression of IRE1a, GADD153, p-PERK, p-elf2a, Caspase7. The aryl hydrocarbon receptor (AhR) and COX-2 is an environmental carcinogen-activated transcription factor associated with tumorigenesis and metastasis. Simultaneously, Eugenol-treated also decreased cooperation of AhR/NF-kB/RelA and COX-2 expression. Similar observations were made when SCM-1, AGS and N87 cells were treated in vitro. Eugeno-induced upregulation of death receptor 5 by increase GADD153(CHOP)/DR5 binding activity but not death receptor 4. Moreover, AhR was down-regulated and cleavaged in the ER fraction of Eugenol-treated cells, as indicated by increased interaction of specific Calpain-10, but not Calpain-1 or Calpain-2. Silence Calpain-10 was abrogated by Eugenol treatment biological effects. In addition, Eugenol inhibited AhR/NF-kB/RelA interaction, nuclear translocation and DNA binding activity in cancer cells in time course and dose-dependent manner by Calpain-10 activation. Taken together, our results suggest that Eugenol suppresses both gastric tumor growth and peritoneal dissemination by inducing apoptosis and activating ER stress.en_US
dc.description.tableofcontents目次 中文摘要....................................................i 英文摘要...................................................ii 目次.....................................................iii 圖目次.....................................................vi 縮寫表....................................................vii 第一章、 前言................................................1 一、 丁香酚(Eugenol) ...................................1 二、 胃癌(Gastric cancer)...............................1 (一) 流行病學...........................................1 1. 遺傳因素...........................................1 2. 性別與年齡.........................................1 3. 飲食生活習慣........................................2 4. 惡性貧血...........................................2 5. 胃部病變及胃切除手術.................................2 6. 幽門螺旋桿菌感染.....................................2 (二) 治療方式...........................................2 三、 COX-2 (Cyclooxygenase-2) .........................3 (一) COX的結構與功能.....................................3 (二) COX-2與癌症........................................3 四、 AhR (aryl hydrocarbon receptor)....................4 (一) AhR與癌症..........................................4 五、 內質網壓力(Endoplasmic Reticulum Stress) ...........4 (一) ER Stress的分子機轉................................4 (二) ER Stress和Calpain................................5 六、 上皮-間質細胞轉換(epithelial-mesenchymal transition 簡稱 EMT).......................................................5 七、 研究方向與動機.......................................6 第二章、 材料與方法...........................................8 一、 實驗儀器............................................8 二、 實驗材料............................................8 (一) 常用緩衝溶液: 附表一.................................8 (二) 實驗試劑: 附表二....................................8 (三) 實驗藥品: 附表三....................................8 (四) 實驗抗體: 附表四....................................8 (五) PCR Primer: 附表五.................................8 三、 實驗方法............................................8 (一) 細胞培養(Cell culture).............................8 (二) 人類臍靜脈內皮細胞初代培養(Primary HUVEC culture).....9 (三) 蛋白質萃取(Protein extraction).....................9 (四) 西方墨點法(Western Blot)...........................9 (五) 核酸萃取(RNA extraction)..........................10 (六) 反轉錄聚合酶連鎖效應(Reverse Transcription-PCR).....10 (七) 聚合酶連鎖效應(Polymerase Chain Reaction)..........10 (八) 免疫螢光染色(Immunofluorescence stain).............11 (九) 免疫組織染色(Immunohistochemistry stain)...........11 (十) 瓊膠生長實驗(Colony Formation Assay)...............11 (十一) 流氏細胞儀分析(Flowcytometry)......................12 (十二) 人類臍靜脈內皮細胞血管新生實驗(HUVEC Tube Formation).........................................12 (十三) 大鼠主動脈環試驗(Rat Aorta Ring Assay)..............12 (十四) 酶譜分析(Gelatin Zymography)......................12 (十五) 酵素活性分析(Enzyme Activity)......................13 (十六) 細胞增生試驗(MTT Assay)............................13 (十七) 細胞核質蛋白分離萃取(Nuclear/ Cytosol protein extraction)........................................13 (十八) 免疫沉澱法(Immunoprecipitation)...................13 (十九) 核酸干擾技術(RNA interference).....................14 (二十) 傷口癒合試驗(Wound Healing Assay)..................14 (二十一) 動物實驗(Animal experiment).......................14 (二十二) 動物組織蛋白萃取....................................15 (二十三) 電泳移動率試驗(Electrophoretic Mobility Shift Assay, EMSA)....................................15 (二十四) 染色質免疫沉澱法(Chromatin Immunoprecipitation, CHIP Assay)......................................15 (二十五) 雞胚胎絨毛膜試驗(Chicken chorioallantoic membrane assays,CAM)..............................................16 (二十六) 彗星試驗(Comet assay)..............................16 (二十七) 分離內質網蛋白(Isolation of ER).....................16 第三章、 實驗結果............................................18 一、 胃癌癌化過程中顯著增加小鼠胃部組織中AhR的蛋白表現 量........................................................18 二、 Eugenol在轉錄層次上透過NFkB及AhR有效抑制胃癌細胞中COX-2的轉錄表現.....................................................18 三、 Eugenol有效抑制NFkB-p65和AhR在胃癌細胞中內生性交 互作用.....................................................18 四、 Eugenol降低AhR在胃癌細胞中的表現......................19 五、 Eugenol誘導胃癌細胞內質網壓力的形成....................19 六、 Eugenol活化胃癌細胞內質網壓力進而活化Calpain的蛋白表現及活性........................................................20 七、 Calpain抑制劑有效恢復胃癌細胞中由Eugenol所減少AhR的表現........................................................20 八、 Eugenol抑制胃癌細胞增生並促進細胞走向死亡...............21 九、 Eugenol促進細胞走向死亡並造成細胞產生DNA碎片............22 十、 以PET/CT影像觀察建立的原位胃癌的情形...................22 十一、 Eugenol抑制胃癌細胞在裸鼠體內生長和擴散之情形............23 十二、 Eugenol有效抑制胃癌細胞在裸鼠皮下及體外試驗之生長........23 十三、 Eugenol有效抑制胃癌細胞的移動性和血管新生作用............24 十四、 Eugenol調控胃癌細胞在上皮-間質細胞轉換(epithelial-mesenchymal transition 簡稱EMT)現象........................25 十五、 總結..............................................26 第四章、討論................................................27 第五章、參考文獻.............................................31 附錄表.....................................................68zh_TW
dc.language.isozh_TWen_US
dc.publisher生物醫學研究所zh_TW
dc.relation.urihttp://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-0107201314491000en_US
dc.subject丁香酚(Eugenol)zh_TW
dc.subjectEugenolen_US
dc.subject芳香烴受體(AHR)zh_TW
dc.subject鈣蛋白酶-10zh_TW
dc.subjectgastric canceren_US
dc.subjectcalpainen_US
dc.subjectperitoneal disseminationen_US
dc.subjectaryl hydrocarbon receptoren_US
dc.title小鼠原位癌模式中丁香酚藉由增加內質網壓力抑制胃癌腫瘤生成以及腹膜轉移zh_TW
dc.titleEugenol suppresses gastric tumor growth and peritoneal dissemination by increasing ER stress in an orthotopic modelen_US
dc.typeThesis and Dissertationzh_TW
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item.languageiso639-1zh_TW-
item.openairetypeThesis and Dissertation-
item.cerifentitytypePublications-
item.openairecristypehttp://purl.org/coar/resource_type/c_18cf-
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